A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus
To establish a rapid and sensitive detection method for the porcine reproductive and respiratory syndrome virus (PRRSV), gene-specific primers and a TaqMan probe were designed based on the <i>M</i> gene of PRRSV, and a new stable fully pre-mixed reverse transcription real-time fluorescen...
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| Format: | Article |
| Language: | English |
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MDPI AG
2025-01-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/12/1/26 |
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| author | Hongri Zhao Xingyu Xiao Yajuan Sun Yang Chen Yongzhe Zhang Peng Li Hui Jin Ying Li Rui Yin |
| author_facet | Hongri Zhao Xingyu Xiao Yajuan Sun Yang Chen Yongzhe Zhang Peng Li Hui Jin Ying Li Rui Yin |
| author_sort | Hongri Zhao |
| collection | DOAJ |
| description | To establish a rapid and sensitive detection method for the porcine reproductive and respiratory syndrome virus (PRRSV), gene-specific primers and a TaqMan probe were designed based on the <i>M</i> gene of PRRSV, and a new stable fully pre-mixed reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) reaction mixture was developed. A simple and rapid RT-qPCR detection method for PRRSV was developed by optimizing nucleic acid amplification conditions. The results showed that the method was able to specifically detect PRRSV without cross-reactivity with the other 11 porcine susceptible viruses. The sensitivities of the assay were 3.12 × 10<sup>0</sup> copies/μL and 10<sup>0</sup> TCID<sub>50</sub>/μL for <i>M</i> gene and virus, respectively, and the repeatability and reproducibility (relative standard deviation, CV) of the assay were less than 2.5%. Based on the new fullly pre-mixed RT-qPCR reaction mixture, the RT-qPCR detection method may provide a new, simple, and rapid method for accurately detecting PRRSV. |
| format | Article |
| id | doaj-art-7a82c5d1b0d34fe88fe9c449bf97ce29 |
| institution | Kabale University |
| issn | 2306-7381 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj-art-7a82c5d1b0d34fe88fe9c449bf97ce292025-01-24T13:52:02ZengMDPI AGVeterinary Sciences2306-73812025-01-011212610.3390/vetsci12010026A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome VirusHongri Zhao0Xingyu Xiao1Yajuan Sun2Yang Chen3Yongzhe Zhang4Peng Li5Hui Jin6Ying Li7Rui Yin8College of Veterinary Medicine, Jilin Agricultural University, Changchun 130118, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaDepartment of Neurology, China-Japan Union Hospital of Jilin University, Changchun 130033, ChinaCollege of Veterinary Medicine, Jilin Agricultural University, Changchun 130118, ChinaCollege of Veterinary Medicine, Jilin Agricultural University, Changchun 130118, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Veterinary Medicine, Jilin Agricultural University, Changchun 130118, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaTo establish a rapid and sensitive detection method for the porcine reproductive and respiratory syndrome virus (PRRSV), gene-specific primers and a TaqMan probe were designed based on the <i>M</i> gene of PRRSV, and a new stable fully pre-mixed reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) reaction mixture was developed. A simple and rapid RT-qPCR detection method for PRRSV was developed by optimizing nucleic acid amplification conditions. The results showed that the method was able to specifically detect PRRSV without cross-reactivity with the other 11 porcine susceptible viruses. The sensitivities of the assay were 3.12 × 10<sup>0</sup> copies/μL and 10<sup>0</sup> TCID<sub>50</sub>/μL for <i>M</i> gene and virus, respectively, and the repeatability and reproducibility (relative standard deviation, CV) of the assay were less than 2.5%. Based on the new fullly pre-mixed RT-qPCR reaction mixture, the RT-qPCR detection method may provide a new, simple, and rapid method for accurately detecting PRRSV.https://www.mdpi.com/2306-7381/12/1/26PRRSV<i>M</i> geneTaqMan RT-qPCRfully pre-mixed reaction mixturedetection |
| spellingShingle | Hongri Zhao Xingyu Xiao Yajuan Sun Yang Chen Yongzhe Zhang Peng Li Hui Jin Ying Li Rui Yin A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus Veterinary Sciences PRRSV <i>M</i> gene TaqMan RT-qPCR fully pre-mixed reaction mixture detection |
| title | A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus |
| title_full | A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus |
| title_fullStr | A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus |
| title_full_unstemmed | A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus |
| title_short | A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus |
| title_sort | simple and sensitive rt qpcr technology for rapid detection of porcine reproductive and respiratory syndrome virus |
| topic | PRRSV <i>M</i> gene TaqMan RT-qPCR fully pre-mixed reaction mixture detection |
| url | https://www.mdpi.com/2306-7381/12/1/26 |
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